Schmitz, C.; Pepelanova, I.; Seliktar, D.; Potekhina, E.; Belousov, V.V. et al.: Live reporting for hypoxia : Hypoxia sensor–modified mesenchymal stem cells as in vitro reporters. In: Biotechnology and Bioengineering 117 (2020), Nr. 11, S. 3265-3276. DOI: https://doi.org/10.1002/bit.27503
Abstract: | |
Natural oxygen gradients occur in tissues of biological organisms and also in the context of three-dimensional (3D) in vitro cultivation. Oxygen diffusion limitation and metabolic oxygen consumption by embedded cells produce areas of hypoxia in the tissue/matrix. However, reliable systems to detect oxygen gradients and cellular response to hypoxia in 3D cell culture systems are still missing. In this study, we developed a system for visualization of oxygen gradients in 3D using human adipose tissue–derived mesenchymal stem cells (hAD-MSCs) modified to stably express a fluorescent genetically engineered hypoxia sensor HRE-dUnaG. Modified cells retained their stem cell characteristics in terms of proliferation and differentiation capacity. The hypoxia-reporter cells were evaluated by fluorescence microscopy and flow cytometry under variable oxygen levels (2.5%, 5%, and 7.5% O2). We demonstrated that reporter hAD-MSCs output is sensitive to different oxygen levels and displays fast decay kinetics after reoxygenation. Additionally, the reporter cells were encapsulated in bulk hydrogels with a variable cell number, to investigate the sensor response in model 3D cell culture applications. The use of hypoxia-reporting cells based on MSCs represents a valuable tool for approaching the genuine in vivo cellular microenvironment and will allow a better understanding of the regenerative potential of AD-MSCs. © 2020 The Authors. Biotechnology and Bioengineering published by Wiley Periodicals LLC | |
License of this version: | CC BY 4.0 Unported |
Document Type: | Article |
Publishing status: | publishedVersion |
Issue Date: | 2020 |
Appears in Collections: | Naturwissenschaftliche Fakultät |
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